Respiratory Virus Research Articles

Seasonality of respiratory, enteric, and urinary viruses revealed by wastewater genomic surveillance.

Wastewater surveillance can reveal population-level infectious disease burden and emergent public health threats can be reliably assessed through wastewater surveillance. While molecular methods for wastewater monitoring of microorganisms have traditionally relied on PCR-based approaches, next-generation sequencing (NGS) can provide deeper insights via genomic analyses of multiple diverse pathogens. We conducted a year-long sequencing surveillance of 1,408 composite wastewater samples collected from 12 neighborhood-level access points in the greater Tempe area, Arizona, USA, and show that variation in wastewater viruses is driven by seasonal time and location. The temporal dynamics of viruses in wastewater were influenced cyclically, with the most dissimilarity between samples 23 weeks apart (i.e., winter vs summer, spring vs fall). We identified diverse urinary and enteric viruses including polyomaviruses, astroviruses, and noroviruses, and showed that their genotypes/subtypes shifted across seasons. We show that while wastewater data of certain respiratory viruses like severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strongly correlate with clinical case rates, laboratory-reported case incidences were discordant with surges of high viral load in wastewater for other viruses like human coronavirus 229E. These results demonstrate the utility of wastewater sequencing for informing decision-making in public health.IMPORTANCEWastewater surveillance can provide insights into the spread of pathogens in communities. Advances in next-generation sequencing (NGS) methodologies allow for more precise detection of viruses in wastewater. Long-term wastewater surveillance of viruses is an important tool for public health preparedness. This system can act as a public health observatory that gives real-time early warning for infectious disease outbreaks and improved response times.

The role of telehealth in the delivery of oral and maxillofacial surgery services at a level 6 hospital in New South Wales Australia during COVID-19 - the John Hunter Hospital experience: a commentary, review and cautionary tale for the future.

The objective of this article is to understand the uptake in use of Telehealth services during the COVID-19 pandemic as a way to provide compelling impetus for further research and promotion of Telehealth in general. A data review of Outpatient Department data from 2020 to 2023, as compiled by the Surgical Outcomes improvement Unit. Telehealth was widely adopted during the COVID-19 pandemic for clinicians to provide healthcare to patients whilst adhering to strict lockdown guidelines aimed at limiting exposure and spread of the respiratory virus. There has been minimal analysis, however, of the ethical and legal implications of telemedicine. Whilst it seems that access to care is improved via telehealth, there are subtle, inherent barriers intrinsic to it. There was a significant increase in the use of audio and audiovisual aids in the delivery of Oral and Maxillofacial Outpatient care during the COVID-19 period. Further analysis and improvements need to be made to the system before it can be considered as sufficient and adequate. Where there is rapid adoption of Telehealth in the delivery of Oral and Maxillofacial Surgery Services, there must be concurrent rigorous review and update of systems in order to address the key areas of patient safety and accuracy of treatment.

PSI-4 Evaluation of chemical mitigants when soybean meal was inoculated with porcine epidemic diarrhea virus, porcine reproductive and respiratory virus, and Seneca Valley virus 1

Abstract Chemical mitigants have been found to decrease virus concentrations in feed and ingredient matrices. Continued research is needed to identify the appropriate inclusion levels and application time for different viruses in these matrices. Therefore, the objective was to evaluate different inclusion levels of two different chemical mitigants when applied either before or after inoculation of porcine epidemic diarrhea virus (PEDV), porcine reproductive and respiratory syndrome virus (PRRSV) and Seneca Valley virus 1 (SVV1) to soybean meal. The experiment was designed in a 2×2×2 factorial with two chemical mitigants, a formaldehyde-based product (Termin-8, Anitox Corp. Lawrenceville, GA) and a product containing Synergistic blend of phytochemicals and carboxylic acids (Finio, Anitox Corp. Lawrenceville, GA), two applications times, before virus inoculation (prevention) and after virus inoculation (mitigation), and either a low or high inclusion level (Termin-8: 2 or 3 kg/MT; Finio: 1.75 or 2.75 kg/MT). On d 1, samples of the respective matrices were weighed in 50 g aliquots and added to 500 mL bottles. Chemical mitigants were applied to the prevention samples at their respective inclusion levels and 50 µL each of 1×107 TCID50/mL PEDV, 1×108 TCID50/mL PRRSV, and 1×108 TCID50/mL SVV1 were added to the mitigation samples. All bottles were shaken and allowed to sit at room temperature for 24 h. On d 2, virus was added to the prevention samples and chemical mitigants were added to the mitigation bottles, samples incubated at room temperature for 1 h before 200 mL of phosphate buffer solution was added, and bottles incubated at 4°C for 18 h. After incubation, liquid from the samples was centrifuged and the supernatant analyzed via triplex PCR at Kansas State University Veterinary Diagnostic Laboratory. Cycle threshold and proportion PCR positive were analyzed using SAS GLIMMIX v 9.4 (SAS, Inc., Cary, NC) with each virus and matrix combination analyzed individually. A chemical mitigant×application time×inclusion level interaction was not observed (P > 0.05) for any virus. A chemical mitigant×application time interaction was observed (P < 0.05) in all three viruses, where less viral RNA was detected (P < 0.05) when either Termin-8 or Finio were applied after inoculation (mitigation) than when applied prior to inoculation or to the positive controls. An application time×inclusion level interaction was noticed (P < 0.05) for SVV1 and PEDV in soybean meal. Chemical mitigants applied at either the low or high inclusion level after inoculation (mitigation) had less detectable RNA (P < 0.05) than their prevention counterparts. Overall, Termin-8 and Finio can reduce detectable RNA immediately in contaminated soybean meal, with ongoing research evaluating the effect of extended contact times between the mitigants and soybean meal.

67 Decontamination of a feed manufacturing environment following inoculation of porcine epidemic diarrhea virus, porcine reproductive and respiratory virus, and Seneca Valley virus 1

Abstract Feed mill decontamination is difficult because equipment is not designed to be cleaned with water. Alternate strategies may improve the ability of a mill to decontaminate in the event of viral contamination. The objective was to evaluate different decontamination strategies within a mill following the inoculation of porcine epidemic diarrhea virus (PEDV), porcine reproductive and respiratory syndrome virus (PRRSV), and Seneca Valley virus 1 (SVV1) contaminated feed. A batch of feed was inoculated with PEDV, PRRSV, and SVV1 and ran through a mixer, bucket elevator, corn cleaner, screw conveyor, distributor head, and spout. Following mill inoculation, decontamination strategies were implemented with environmental samples collected after each decontamination step. Decontamination strategies included: 1) complete facility decontamination including organic matter removal with hot water power washing, disinfection with 1% peroxygen (Virkon S, Lanxess, Germany) followed by a rinse, and a second disinfection with 5% sodium hypochlorite (Clorox, Oakland, CA) followed by a heating period for 48 h once the facility reached 60°C; 2) chlorine dioxide application (ProOxine AH, Bio-Cide International, Inc., Norman, OK); 3) organic matter removal using vacuums (Ridge Tool Company, Elyria, OH) and chlorine dioxide application; 4) heat up with portable electric heaters for exactly 48 h; and 5) organic matter removal and heat up with portable heaters for exactly 48 h. Samples were analyzed via triplex PCR. Cycle threshold and proportion PCR positive were analyzed using SAS GLIMMIX v 9.4 (SAS, Inc., Cary, NC). A swine bioassay was completed to determine the infectivity of samples following the final decontamination step of each treatment. A treatment × decontamination step × location interaction was observed (P < 0.05) for SVV1, where less RNA was detected post-treatment compared with pre-treatment following complete facility decontamination on surfaces including the mixer, corn cleaner, screw conveyor, and floor around the feed discharge (P < 0.05). There was no statistical difference between pre- and post-treatment samples for other combinations of decontamination treatment and location (P > 0.05), although numerical increases in Ct were generally observed following treatment. Across all treatments, the act of decontamination reduced detectable PEDV (P < 0.05) and PRRSV (P < 0.05) RNA when compared with samples immediately following inoculation, but complete facility decontamination was the only treatment where PEDV and PRRSV was non-detectable in all locations. Pigs given samples post-treatment showed no evidence of SVV1 or PEDV infection. Although PRRSV RNA was rarely found via PCR from samples collected within the mill, PRRSV infection was observed in pigs given the chlorine dioxide with and without organic matter removal treatments and the organic matter removal plus heat treatment. Overall, all treatments reduced detectable RNA for all viruses between the inoculation step and the final decontamination step; however, PRRSV particles remained infectious following decontamination regardless of PCR results in a swine bioassay.

PSV-18 Improved performance of PRRSV challenged pigs fed 1-α-glycerol monolaurate

Abstract Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Porcine Enteric Diarrhea (PEDV) has posed a persistent challenge to the swine industry despite vaccinations and biosecurity measures. Feed has been shown to be a carrier for bacterial and viral pathogens (Dee et al., 2020). Feed mitigation using medium chain fatty acids can eliminate the pathogen or reduce the pathogen load below its infectious dose. However, less is known regarding the application of monoglycerides for feed mitigation. The objective of this study was to determine if using only 1-α-glycerol monolaurate (GML) can be an effective feed mitigant against PRRSV compared with untreated control feed. A total of 2 rooms, each with 100 pigs per room, and 6 pens per room were used. The initial body weight of the pigs averaged 15 kg and were sourced from a herd free of PRRSV and PEDV. GML was added to the feed at 2 kg/mT. The study period was 14 d with starting and end body weights measured. An ice block viral challenge as described by Dee et al. (2020) was used. Briefly, a 500 mL ice cube containing 1x107 50% tissue culture infectious dose of each virus [PRRSV, PEDV and Senecavirus A (SVA)] was added to 1,300 kg of feed on d 0 and again on d 6. On d 15 pigs were tested for PRRSV (serum), PEDV (rectal swabs) and SVA (tonsil samples). Control and GML fed pigs did not seroconvert to PEDV and SVA, therefore, only results for PRRSV are presented. Average daily gain was significantly improved in GML fed pigs (0.50 kg/d vs 0.20 kg/d; P < 0.05). Percentage of PRRSV positive feed samples were decreased (P < 0.05) in GML-treated feed from 100% (Ct = 32.2) in control feed to 33% (Ct = 34.3). The percentage of positive oral fluid samples decreased (P < 0.05) from 100% (Ct = 26.1) in control pigs to 17.0% (Ct = 33.0) in GML fed pigs. GML fed pigs exhibited no clinical signs of PRRS (dysponea), whereas 100% of the control pigs had clinical signs (P < 0.05). Similarly, GML fed pigs had no PRRSV positive serum samples but 100% of the control pigs had PRRSV positive serum samples. Results from this study show that the use of only GML can be an effective feed mitigant to reduce or eliminant the clinical signs of PRRS in pigs and maintain growth performance.

PSIII-20 Impact of EnduraPig on Performance of PRRSV-Negative growing-finishing pigs

Abstract A total of 479 weaned pigs [PIC Camborough × PIC 337; initial body weight (BW) = 26.8 kg; Porcine reproductive and respiratory syndrome virus, PRRSV-negative] were used to evaluate the effects of EnduraPig (PMI Additives), a patent-pending feed additive that has been designed to support health, performance, and immune function of pigs, on growth performance of growing-finishing pigs. Pigs were blocked by initial BW and gender and assigned to 48 pens, which were allotted to one of 4 dietary treatments (12 pens/treatment, 9-10 pigs/pen), including T1) Control, T2) EnduraPig 0.1% (EP low), T3) EnduraPig 0.2% (EP medium), and T4) EnduraPig 0.3% (EP high). The EnduraPig inclusion levels were reduced by one-half when pigs reached approximately 70 kg BW (d 42). A five-phase feeding program was used with all diets formulated on a corn-DDGS-soybean meal basis to meet the nutrient requirements of growing-finishing pigs. Analysis of variance for growth measures was performed using the MIXED Procedure of SAS (SAS 9.4), and polynomial contrasts were used to evaluate linear, quadratic, and cubic effects of the EP levels. Significant differences were declared at P ≤ 0.05 and trends at P ≤ 0.10. During d 0-72, feeding EnduraPig tended to improve average daily gain (ADG) (0.96, 0.97, 1.00, and 0.97 kg, respectively; cubic, P = 0.06) with the greatest ADG observed in EP medium. During d 72 to market, feeding EnduraPig increased ADG (1.13, 1.14, 1.18, and 1.17 kg, respectively; linear, P = 0.02), average daily feed intake (ADFI; 3.48, 3.42, 3.55, and 3.49, respectively; cubic, P = 0.02) and feed:gain (3.08, 3.00, 3.01, and 2.98, respectively; linear, P = 0.10). Overall (d 0-market), feeding EnduraPig enhanced ADG (1.02, 1.02, 1.05, and 1.03 kg, respectively; linear, quadratic, and cubic, P = 0.04, 0.09, 0.01), with the greatest ADG observed in EP medium resulting in a 4.0 kg greater market BW compared with the control (128.9, 129.2, 132.9, and 130.5, respectively; linear and cubic, P < 0.05). In addition, feeding EnduraPig numerically improved feed:gain (2.51, 2.48, 2.48, and 2.48, respectively; linear, P = 0.14). With respect to BW variation, feeding EnduraPig reduced the standard deviation in market BW (10.65, 8.06, 8.86, and 9.54 kg, respectively; quadratic, P < 0.05), with the greatest reduction from EP low and EP medium (by 24 and 17%, respectively) compared with the control. In summary, data from this study indicate that feeding EnduraPig at 0.2% from 27 kg to 70 kg BW and 0.1% from 70 to market improves growth performance and reduces BW variation of PRRSV-negative growing-finishing pigs.

PSIII-19 Impact of EnduraPig and EnduraSow supplementation on growth performance of wean to finish pigs naturally infected with porcine reproductive and respiratory syndrome virus

Abstract Porcine reproductive and respiratory syndrome (PRRS) continues to be an economically devastating disease to the pork industry around the world. Apart from vaccination and biosecurity measures, nutritional strategies are needed to mitigate the negative effects of PRRS virus (PRRSV) infection on pork production. The aim of the present study was to determine the effect of EnduraPig and EnduraSow (PMI Additives) on growth performance, viremia, and livability of wean to finish pigs naturally infected with PRRSV. A total of 660 newly-weaned pigs [DNA L600 x 241 genetics; average body weight (BW) = 7.3 ± 0.14 kg) were divided into 33 pens (11 pens per treatment, 20 pigs per pen). Each pen within a location block was randomly assigned to one of three dietary treatments consisting of either a non-supplemented basal diet (NC), basal diet with EnduraPig (EP), or basal diet with EnduraSow (ES). Apart from growth [average daily gain (ADG), average daily feed intake (ADFI), feed to gain ratio (F:G)] and livability metrics, serum sample was also collected from 1 sentinel pig per pen at 0, 21, 42, and 63 d postweaning to monitor recovery from PRRSV infection, based on the change in percentage of viremic pigs. Analysis of variance for growth and mortality was performed using Mixed Procedure in SAS (SAS 9.4, SAS Institute Inc., Cary, NC) and a priori contrasts were used to detect treatment differences at P < 0.05. Association between treatments and % viremic pigs at each time point was assessed using Fisher’s exact test and declared significant at P < 0.05. Mortality and removals were greatest during the first 4 wk postweaning, but no treatment differences were detected in overall wean to finish mortality (23.6, 21.8, and 20.4% for NC, EP, and ES, respectively). At d 21, 89.9% of the pigs sampled were viremic compared with 56.8% on d 0. The average % viremic pigs went down to 57.5 and 18.5%, for d 42 and 63, respectively, with no treatment differences observed in both periods. Feeding EP, however, improved all growth performance metrics compared with NC (+11.8 kg in final body weight, P < 0.01; +13% in ADG, P < 0.01; +7% in ADFI, P < 0.05; and -5.5% in F:G, P < 0.05). Feeding ES also improved performance of wean to finish pigs over NC (+16.4 kg in final body weight, P < 0.01; +18% in ADG, P < 0.01; +12% in ADFI, P < 0.01; and -5.2% in F:G, P < 0.05). In summary, the results of this study indicate that dietary supplementation of EnduraPig or EnduraSow improved growth performance of wean to finish pigs naturally infected with PRRSV, indicating that these nutritional technologies help boost the immune response against PRRSV.

208 The antiviral and immunomodulating properties from glycerides of lauric acid in PRRSV-infected PAM cells

Abstract The objective of this study was to assess the ability of a commercially available blend of glycerides of C12 to inhibit porcine reproductive and respiratory syndrome virus (PRRSV) replication in porcine alveolar macrophages (PAM) and to investigate the immunomodulatory effect during PRRSV infection in vitro. PAM were collected by lung lavage from 4-wk-old piglets, free of PRRSV. PRRSV strain 13V091 was used in this study and a commercially available mixture of mono-, di- and tri-glycerides of lauric acid (GC12). The product concentrations tested were 0.022% (T1), 0.044% (T2) and 0.088% (T3). Three different infection assays were used: (1) Co-incubation; PRRSV was inoculated together with GC12 for 1 h, (2) Pre- and Post-infection; GC12 was added to PAM cells for 24 h before infection and after 1 h of incubation for another 24 h, and (3) Post-infection; GC12 was added only after PRRSV infection for 24 h. Titration of the progeny virus was performed on PAM from all three infection assays. Cytokine analysis was only performed for infection assay 2 and 3. Collected supernatants were subjected to multiplex ELISA to measure the level of TNF-α and IL-8. Two-tailed t test P value was used to determine statistically significant differences. P < 0.05 was considered to be statistically significant. Addition of GC12 in the Co-incubation assay did not reduce PRRSV infectivity at 24-h post infection (hpi; Table 1). In contrast, the Pre- and Post-infection assay showed significantly reduced PRRSV titer from 6 hpi onwards at concentration 0.044%. At concentration 0.088% a significant reduction was observed at 24 hpi. Addition of GC12 after PRRSV inoculation (post-infection) also showed significantly reduced PRRSV titer from 6 hpi onwards at concentration 0.044% and 0.088%. In the Pre- and Post-infection assay, all treatments with GC12 numerically increased the production of TNF-α at 24 hpi. A dose response effect was observed at 24 hpi with increasing concentrations of GC12 resulting in increasing levels of TNF-α. However, in the Post-infection assay only treatment with 0.088% GC12 numerically increased the production of TNF-α by 1.5-fold. Compared with the control (PAM+PRRSV), a significantly greater concentration of IL-8 was observed with 0.044% (P = 0.019) and 0.088% (P = 0.007) GC12 24 hpi in the Pre- and Post-infection assay (1,806 ± 376; 3,751 ± 636; 3,439 ± 420 pg/mL respectively). These two concentrations resulted in numeric increase of IL-8 24 hpi in the Post-infection assay and no effect was observed with 0.022% GC12 (Control 2,332 ± 998; T1 2,079 ± 964; T2 3,525 ± 344; T3 4,475 ± 295). From this study it can be concluded that GC12 exhibited antiviral activity against PRRSV strain 13V091 and influences the cytokine production of PAM during PRRSV infection, potentially neutralizing the suppression of the immune response by PRRSV.

Development and application of a high-sensitivity immunochromatographic test strip for detecting pseudorabies virus

IntroductionPseudorabies (PR) is a multi-animal comorbid disease caused by pseudorabies virus (PRV), which are naturally found in pigs. At the end of 2011, the emergence of PRV variant strains in many provinces in China had caused huge economic losses to pig farms. Rapid detection diagnosis of pigs infected with the PRV variant helps prevent outbreaks of PR. The immunochromatography test strip with colloidal gold nanoparticles is often used in clinical testing due to its low cost and high throughput.MethodsThis study was designed to produce monoclonal antibodies targeting PRV through immunization of mice using the eukaryotic system to express the gE glycoprotein. Subsequently, paired monoclonal antibodies were screened based on their sensitivity and specificity for use in the preparation of test strips.Results and discussionThe strip prepared in this study was highly specific, only PRV was detected, and there was no cross-reactivity with glycoprotein gB, glycoprotein gC, glycoprotein gD, and glycoprotein gE of herpes simplex virus and varicellazoster virus, porcine epidemic diarrhea virus, Senecavirus A, classical swine fever virus, porcine reproductive and respiratory syndrome virus, and porcine parvovirus. Moreover, it demonstrated high sensitivity with a detection limit of 1.336 × 103 copies/μL (the number of viral genome copies per microliter); the coincidence rate with the RT-PCR detection method was 96.4%. The strip developed by our laboratory provides an effective method for monitoring PRV infection and controlling of PR vaccine quality.

Adverse outcomes in patients hospitalized with pneumonia at age 60 or more: A prospective multi-centric hospital-based study in India.

Limited data exists regarding risk factors for adverse outcomes in older adults hospitalized with Community-Acquired Pneumonia (CAP) in low- and middle-income countries such as India. This multisite study aimed to assess outcomes and associated risk factors among adults aged ≥60 years hospitalized with pneumonia. Between December 2018 and March 2020, we enrolled ≥60-year-old adults admitted within 48 hours for CAP treatment across 16 public and private facilities in four sites. Clinical data and nasal/oropharyngeal specimens were collected by trained nurses and tested for influenza, respiratory syncytial virus (RSV), and other respiratory viruses (ORV) using the qPCR. Participants were evaluated regularly until discharge, as well as on the 7th and 30th days post-discharge. Outcomes included ICU admission and in-hospital or 30-day post-discharge mortality. A hierarchical framework for multivariable logistic regression and Cox proportional hazard models identified risk factors (e.g., demographics, clinical features, etiologic agents) associated with critical care or death. Of 1,090 CAP patients, the median age was 69 years; 38.4% were female. Influenza viruses were detected in 12.3%, RSV in 2.2%, and ORV in 6.3% of participants. Critical care was required for 39.4%, with 9.9% in-hospital mortality and 5% 30-day post-discharge mortality. Only 41% of influenza CAP patients received antiviral treatment. Admission factors independently associated with ICU admission included respiratory rate >30/min, blood urea nitrogen>19mg/dl, altered sensorium, anemia, oxygen saturation <90%, prior cardiovascular diseases, chronic respiratory diseases, and private hospital admission. Diabetes, anemia, low oxygen saturation at admission, ICU admission, and mechanical ventilation were associated with 30-day mortality. High ICU admission and 30-day mortality rates were observed among older adults with pneumonia, with a significant proportion linked to influenza and RSV infections. Comprehensive guidelines for CAP prevention and management in older adults are needed, especially with the co-circulation of SARS-CoV-2.

Fiber rich food suppressed airway inflammation, GATA3 + Th2 cells, and FcεRIα+ eosinophils in asthma

BackgroundAllergic Asthma is a disease presenting various endotypes and no current therapies act curative but alleviate disease symptoms. Dietary interventions are gaining increasing importance in regulating immune responses. Furthermore, short chain fatty acids (SFCA), as the main products of dietary fiber’s fermentation by the gut bacteria, ameliorate the pathogenesis and disease burden of different illnesses including asthma. Nevertheless, the connection and crosstalk between the gut and lung is poorly understood.ObjectiveIn this work, the role of high fiber diet on the development of allergic asthma at baseline and after exacerbation of disease induced by respiratory viruses was investigated.MethodsHereby, SCFA in serum of asthmatic and non-asthmatic pre-school children before and after airway disease symptoms were analyzed. Moreover, the effect of high fiber diet in vivo in a murine model of house dust mite extract (HDM) induced allergic asthma and in the end in isolated lung and spleen cells infected ex vivo with Rhinovirus was analyzed.ResultsIn this study, a decrease of the SCFA 3-Hydroxybutyric acid in serum of asthmatic children after symptomatic episodes at convalescent visit as compared to asthmatic and control children at baseline visit was observed. In experimental asthma, in mice fed with high fiber diet, a reduced lung GATA3 + Th2 type mediated inflammation, mucus production and collagen deposition and expression of Fc epsilon receptor Ia (FcεRIa) in eosinophils was observed. By contrast, the CD8+ memory effector T cells were induced in the lungs of asthmatic mice fed with high fiber diet. Then, total lung cells from these asthmatic mice fed with either standard food or with fiber rich food were infected with RV ex vivo. Here, RV1b mRNA was found significantly reduced in the lung cells derived from fiber rich food fed mice as compared to those derived from standard food fed asthmatic mice. Looking for the mechanism, an increase in CD8+ T cells in RV infected spleen cells derived from fiber rich fed asthmatic mice, was observed.ConclusionConvalescent preschool asthmatic children after a symptomatic episode have less serum ß-Hydroxybutyric acid as compared to control and asthmatic children at baseline visit. Fiber rich diet associated with anti-inflammatory effects as well as anti-allergic effects by decreasing Type 2 and IgE mediated immune responses and inducing CD8+ memory effector T cells in a murine model of allergic asthma. Finally, ex vivo infection with Rhinovirus (RV) of total lung cells from asthmatic mice fed with fiber rich food led to a decreased RV load as compared to mice fed with standard food. Moreover, spleen cells derived from asthmatic mice fed with fiber rich food induced CD8+ T cells after ex vivo infection with RV.Clinical implicationsDietary interventions with increased content in natural fibers like pectins would ameliorate asthma exacerbations. Moreover, respiratory infection in asthma downregulated SCFA in the gut contributing to asthma exacerbations.

ADLM Guidance Document on Laboratory Diagnosis of Respiratory Viruses.

Respiratory viral infections are among the most frequent infections experienced worldwide. The COVID-19 pandemic has highlighted the need for testing and currently several tests are available for the detection of a wide range of viruses. These tests vary widely in terms of the number of viral pathogens included, viral markers targeted, regulatory status, and turnaround time to results, as well as their analytical and clinical performance. Given these many variables, selection and interpretation of testing requires thoughtful consideration. The current guidance document is the authors' expert opinion based on the preponderance of available evidence to address key questions related to best practices for laboratory diagnosis of respiratory viral infections including who to test, when to test, and what tests to use. An algorithm is proposed to help laboratories decide on the most appropriate tests to use for the diagnosis of respiratory viral infections.

Validation of an automated, end-to-end metagenomic sequencing assay for agnostic detection of respiratory viruses.

Current molecular diagnostics are limited in the number and type of detectable pathogens. Metagenomic next generation sequencing (mNGS) is an emerging, and increasingly feasible, pathogen-agnostic diagnostic approach. Translational barriers prohibit the widespread adoption of this technology in clinical laboratories. We validate an end-to-end mNGS assay for detection of respiratory viruses. Our assay is optimized to reduce turnaround time, lower cost-per-sample, increase throughput, and deploy secure and actionable bioinformatic results. We validated our assay using residual nasopharyngeal swab specimens from Vancouver General Hospital (n = 359), RT-PCR-positive, or negative for Influenza, SARS-CoV-2, and RSV. We quantified sample stability, assay precision, the effect of background nucleic acid levels, and analytical limits of detection. Diagnostic performance metrics were estimated. We report that our mNGS assay is highly precise, semi-quantitative, with analytical limits of detection ranging from 103-104 copies/mL. Our assay is highly specific (100%) and sensitive (61.9% Overall: 86.8%; RT-PCR Ct < 30). Multiplexing capabilities enable processing of up to 55-specimens simultaneously on an Oxford Nanopore GridION device, with results reported within 12-hours. This study outlines the diagnostic performance and feasibility of mNGS for respiratory viral diagnostics, infection control, and public health surveillance. We addressed translational barriers to widespread mNGS adoption.

Highly Pathogenic Avian Influenza (HPAI) H5 Clade 2.3.4.4b Virus Infection in Birds and Mammals

Avian influenza viruses (AIVs) are highly contagious respiratory viruses of birds, leading to significant morbidity and mortality globally and causing substantial economic losses to the poultry industry and agriculture. Since their first isolation in 2013-2014, the Asian-origin H5 highly pathogenic avian influenza viruses (HPAI) of clade 2.3.4.4b have undergone unprecedented evolution and reassortment of internal gene segments. In just a few years, it supplanted other AIV clades, and now it is widespread in the wild migratory waterfowl, spreading to Asia, Europe, Africa, and the Americas. Wild waterfowl, the natural reservoir of LPAIVs and generally more resistant to the disease, also manifested high morbidity and mortality with HPAIV clade 2.3.4.4b. This clade also caused overt clinical signs and mass mortality in a variety of avian and mammalian species never reported before, such as raptors, seabirds, sealions, foxes, and others. Most notably, the recent outbreaks in dairy cattle were associated with the emergence of a few critical mutations related to mammalian adaptation, raising concerns about the possibility of jumping species and acquisition of sustained human-to-human transmission. The main clinical signs and anatomopathological findings associated with clade 2.3.4.4b virus infection in birds and non-human mammals are hereby summarized.

Drugs targeting CMPK2 inhibit pyroptosis to alleviate severe pneumonia caused by multiple respiratory viruses.

Severe pneumonia caused by respiratory viruses has become a major threat to humans, especially with the SARS-CoV-2 outbreak and epidemic. The aim of this study was to investigate the universal molecular mechanism of severe pneumonia induced by multiple respiratory viruses and to search for therapeutic strategies targeting this universal molecular mechanism. The common differential genes of four respiratory viruses, including respiratory syncytial virus (RSV), rhinovirus, influenza, and SARS-CoV-2, were screened by GEO database, and the hub gene was obtained by Sytohubba in Cytoscape. Then, the effect of hub genes on inflammasome and pyrodeath was investigated in the model of RSV infection in vitro and in vivo. Finally, through virtual screening, drugs targeting the hub gene were obtained, which could alleviate severe viral pneumonia in vitro and in vivo. The results showed that CMPK2 is one of the hub genes after infection by four respiratory viruses. CMPK2 activates the inflammasome by activating NLRP3, and promotes the releases of inflammatory factors interleukin (IL)-1β and IL-18 to induce severe viral pneumonia. Z25 and Z08 can reduce the expression level of CMPK2 mRNA and protein, thereby inhibiting NLRP3 and alleviating the development of severe viral pneumonia. In conclusion, the inflammatory response mediated by CMPK2 is the common molecular mechanism of severe pneumonia induced by viral infection, and Z25 and Z08 can effectively alleviate viral infection and severe pneumonia through this mechanism.

Assessment of Illness Severity in Adults Hospitalized With Acute Respiratory Tract Infection due to Influenza, Respiratory Syncytial Virus, or Human Metapneumovirus.

Influenza, respiratory syncytial virus (RSV), and human metapneumovirus (hMPV) are common respiratory viruses causing similar symptoms. Optimal tools to assess illness severity for these viruses have not been defined. Using the Hospitalized Acute Respiratory Tract Infection (HARTI) study data, we report symptom severity by clinician-rated clinical severity scores (CSS) in adults with influenza, RSV, or hMPV and correlations between CSS and patient-reported outcomes (PROs). HARTI was a global epidemiologic study in adults hospitalized with acute respiratory tract infections. Patients were assessed at enrollment within 24 h of admission with CSS and twice during hospitalization with CSS, Respiratory Infection Intensity and Impact Questionnaire™ (RiiQ™), and EQ-5D-5L. Data were summarized descriptively, stratified by pathogen and baseline and hospitalization characteristics. Domain (general, upper respiratory, and lower respiratory) and sign/symptom subscores are presented for CSS; sign/symptom subscores are presented for RiiQ™ results. Data from 635 patients with influenza, 248 with RSV, and 107 with hMPV were included. At enrollment, total CSS and general and lower respiratory signs/symptoms (LRS) scores were higher for RSV and hMPV than influenza. Between-pathogen differences were greatest for LRS scores. Dyspnea, rales/rhonchi, wheezing, and shortness of breath scores trended higher for RSV and hMPV than influenza. RiiQ™ scores for cough, fatigue, and short of breath were strongly correlated with corresponding clinician-rated symptoms. These findings support the use of PROs (e.g., the RiiQ™) correlating with clinician assessments to gauge patient well-being and aid patient management by accurately assessing respiratory illness severity due to RSV, hMPV, or influenza.

Novel insights into the host cell glycan binding profile of human metapneumovirus.

Numerous viruses have been found to exploit glycoconjugates expressed on human cells as their initial attachment factor for viral entry and infection. The virus-cell glycointeractome, when characterized, may serve as a template for antiviral drug design. Heparan sulfate proteoglycans extensively decorate the human cell surface and were previously described as a primary receptor for human metapneumovirus (HMPV). After respiratory syncytial virus, HMPV is the second most prevalent respiratory pathogen causing respiratory tract infection in young children. To date, there is neither vaccine nor drug available to prevent or treat HMPV infection. Using a multidisciplinary approach, we report for the first time the glycointeractome of the HMPV fusion (F) protein, a viral surface glycoprotein that is essential for target-cell recognition, attachment, and entry. Our glycan microarray and surface plasmon resonance results suggest that Galβ1-3/4GlcNAc moieties that may be sialylated or fucosylated are readily recognized by HMPV F. The bound motifs are highly similar to the N-linked and O-linked glycans primarily expressed on the human lung epithelium. We demonstrate that the identified glycans have the potential to compete with the cellular receptors used for HMPV entry and consequently block HMPV infection. We found that lacto-N-neotetraose demonstrated the strongest HMPV binding inhibition in a cell infection assay. Our current findings offer an encouraging and novel avenue for the design of anti-HMPV drug candidates using oligosaccharide templates.IMPORTANCEAll cells are decorated with a dense coat of sugars that makes a sugar code. Many respiratory viruses exploit this sugar code by binding to these sugars to cause infection. Human metapneumovirus is a leading cause for acute respiratory tract infections. Despite its medical importance, there is no vaccine or antiviral drug available to prevent or treat human metapneumovirus infection. This study investigates how human metapneumovirus binds to sugars in order to more efficiently infect the human host. We found that human metapneumovirus binds to a diverse range of sugars and demonstrated that these sugars can ultimately block viral infection. Understanding how viruses can take advantage of the sugar code on our cells could identify new intervention and treatment strategies to combat viral disease.

Transition from one commercial porcine reproductive and respiratory syndrome modified-live virus vaccine to another in a breeding herd and impact on productivity

Porcine reproductive and respiratory syndrome (PRRS) continues to represent a significant cost to the swine industry and efforts are focused on prevention and mitigation of losses across production phases. Herein describes a PRRS modified-live virus (MLV) vaccinated breeding herd that changed commercial MLV vaccines to improve post-weaning performance. Two whole-herd vaccinations with a new PRRS MLV vaccine, administered 4 weeks apart, occurred without breeding herd production disruptions and with limited changes in diagnostic results. Replacement gilts tested PRRS virus negative 10 weeks post vaccination with the new MLV vaccine. Diagnostics were intermittently positive in the breeding herd and early nursery.

Porcine circovirus 2d associated with stillborn mummified foetus of pig

Recurrent occurrence of stillborn piglets was noticed along with live births during farrowing in pigs in an organized pig farm of Uttar Pradesh. On detailed post mortem examination, externally, the foetus was found to be partially dehydrated and dark brown in colour with shrunken eyes. Internally, visceral organs including lungs, heart, kidneys, liver, spleen and intestine were partially dehydrated and dark brown in colour. Upon molecular investigation, the pooled tissue samples were found positive for porcine circovirus 2 (PCV2) infection using PCR technique and further confirmed the PCV2d genotype by DNA sequencing. Other most possible infections including porcine parvoviral infection (PPV), PCV3 and porcine respiratory and reproductive syndrome virus infection (PRRS) were found negative using PCR and RT-PCR techniques, respectively. This result is suggestive of role of PCV2d as a possible etiological agent to cause stillbirth and mummification of porcine foetuses.

Respiratory Virus Panel Testing in Pediatric Emergency Medicine: An Institutional Experience from 2019 to 2022

Respiratory Virus Panel Testing in Pediatric Emergency Medicine: An Institutional Experience from 2019 to 2022